Nayernia K, von Mering M H, Kraszucka K, Burfeind P, Wehrend A, Köhler M, Schmid M, Engel W
Institute of Human Genetics, University of Göttingern, Germany.
Biol Reprod. 1999 Jun;60(6):1488-95. doi: 10.1095/biolreprod60.6.1488.
The successful production of male gametes requires extensive and precise interactions between germ cells and surrounding testicular somatic cells. We have made use of the mRNA differential display technique to isolate genes involved in germ cell-Sertoli cell interaction. We have identified five differential cDNA bands by comparing RNA from Sertoli cells, spermatids and spermatid-Sertoli cell cocultures. One of the isolated cDNA fragments detected a 1. 4-kilobase (kb) testis- and spermatid-specific transcript (designated as THEG: testicular haploid expressed gene). Northern blot analysis on RNA from spermatids and spermatid-Sertoli cell cocultures demonstrated that Sertoli cells are required for the continued expression of THEG in spermatids. We found two alternatively spliced transcripts for the THEG gene with 1437 base pairs (bp) and 1375 bp by using reverse transcription-polymerase chain reaction. The two open reading frames of 376 amino acids and 181 amino acids coded for putative nuclear proteins. The gene is approximately 10 kb pairs in size, contains 8 exons, and was mapped on mouse chromosome 10 to region B5-C1. Comparison of the two cDNA sequences with the genomic sequence indicated that the smaller transcript lacks exon 4. The differential gene expression of THEG in spermatid-Sertoli cell coculture supports the relevance of germ cell-Sertoli cell interaction for gene regulation during spermatogenesis.
雄性配子的成功产生需要生殖细胞与周围睾丸体细胞之间广泛而精确的相互作用。我们利用mRNA差异显示技术来分离参与生殖细胞-支持细胞相互作用的基因。通过比较支持细胞、精子细胞以及精子细胞-支持细胞共培养物中的RNA,我们鉴定出了五条差异cDNA条带。其中一个分离出的cDNA片段检测到一个1.4千碱基(kb)的睾丸和精子细胞特异性转录本(命名为THEG:睾丸单倍体表达基因)。对精子细胞和精子细胞-支持细胞共培养物中的RNA进行Northern印迹分析表明,支持细胞是精子细胞中THEG持续表达所必需的。通过逆转录-聚合酶链反应,我们发现THEG基因有两个选择性剪接的转录本,分别为1437个碱基对(bp)和1375 bp。这两个开放阅读框分别编码376个氨基酸和181个氨基酸的假定核蛋白。该基因大小约为10 kb对,包含8个外显子,并定位于小鼠染色体10的B5-C1区域。将两个cDNA序列与基因组序列进行比较表明,较小的转录本缺少外显子4。THEG在精子细胞-支持细胞共培养物中的差异基因表达支持了生殖细胞-支持细胞相互作用在精子发生过程中对基因调控的相关性。